( 4 A quantitative analysi...)Versatile Transfection Reagent Offers Low Toxicity and Consistent,,,Performance

A quantitative analysis of transfection efficiencies is shown for LipofectAMINE reagent (Figure 4A) and LipoTAXI reagent (Figure 4B). For all tested conditions, LipoTAXI transfection reagent yielded higher transfection efficiencies of SW13 cells with the pEGFP-C3 plasmid. In addition, cell death, indicated by floating cells, occurred with greater frequency with increasing concentrations of LipofectAMINE reagent. Increased cell death was not seen with increasing concentrations of LipoTAXI reagent, again demonstrating LipoTAXIs low toxicity.

Conclusions

Optimizing transfection conditions is a cell-line dependent process and should be performed for each cell line studied. When comparing efficiencies using four different lipid reagents for transfections in A549, COS-7, and NIH 3T3 cells, Stratagenes LipoTAXI transfection reagent consistently gave the highest transfection efficiencies and the lowest cellular toxicity. Atypical optimization profiles and low transfection efficiencies can be caused by mycoplasma contamination of cell lines, which is easily detected with Stratagenes Mycoplasma PCR Primer Set. LipoTAXI transfection reagent is significantly more effective than LipofectAMINE reagent for transfecting enhanced green fluorescent protein into SW13 cells, as measured by transfection efficiency and cell death. Furthermore, LipoTAXI transfection reagent is effective over a broader range of conditions than LipofectAMINE reagent leading to less cell death. Therefore, LipoTAXI transfection reagent is superior for consistent, low-toxicity, high-efficiency transfections.

REFERENCES

1. Ausubel, F.M., et al. (1994) In Current Protocols in Molecular Biology. Wiley & Sons, New York.

2. Jordan, M., Schallhorn, A., and
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